Using micropatterned lipid bilayer arrays to measure the effect of membrane composition on merocyanine 540 binding.
Using micropatterned lipid bilayer arrays to measure the effect of membrane composition on merocyanine 540 binding.
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使用微图案脂质双层阵列测量膜成分对部花青 540 结合的影响。
DOI：
10.1016/j.bbamem.2011.02.017
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Conboy,JohnC
中科院分区：
文献类型：
--
作者：
Smith,KathrynA;Conboy,JohnC
以亲脂性染料花菁540(MC540)为模型，利用三维连续流动显微镜(CFM)制备了微图案化脂质双层阵列(MLBA)。利用荧光显微镜同时监测MC540与15种不同双层成分的结合情况。对于由液晶(L.C.)组成的双层膜，MC540的荧光强度要大两倍。与凝胶相的双分子膜(1，2-二棕榈酰甘油-3-磷胆碱和1，2-二硬脂酰甘油-3-磷胆碱(DOPC)，1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine)进行了比较。胆固醇(CHO)对MC540与膜结合的影响依赖于膜上的脂质成分，胆固醇降低了DMPC、DPPC和DSPC双层膜与MC540的结合量，而对其余的膜结合率几乎没有影响。相态脂类。当1，2-二油酰基-n-甘油-3-磷酸-L-丝氨酸(钠盐)引入DOPC双层膜时，MC540的荧光强度也有所降低。表面电荷密度的增加似乎减少了强荧光单体的出现，并通过静电斥力增加了弱荧光二聚体的形成。本文证明了MLBA是制备高密度可重复的双层阵列的有用工具，可以高通量地研究小分子-膜相互作用。
The lipophilic dye merocyanine 540 (MC540) was used to model small molecule–membrane interactions using micropatterned lipid bilayer arrays (MLBAs) prepared using a 3D Continuous Flow Microspotter (CFM). Fluorescence microscopy was used to monitor MC540 binding to fifteen different bilayer compositions simultaneously. MC540 fluorescence was two times greater for bilayers composed of liquid-crystalline (l.c.) phase lipids (1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC),1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)) compared to bilayers in the gel phase (1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC)). The effect cholesterol (CHO) had on MC540 binding to the membrane was found to be dependent on the lipid component; cholesterol decreased MC540 binding in DMPC, DPPC and DSPC bilayers while having little to no effect on the remaining l.c. phase lipids. MC540 fluorescence was also lowered when 1,2-dioleoyl-sn-glycero-3-phospho-l-serine (sodium salt) (DOPS) was incorporated into DOPC bilayers. The increase in the surface charge density appears to decrease the occurrence of highly fluorescent monomers and increase the formation of weakly fluorescent dimers via electrostatic repulsion. This paper demonstrates that MLBAs are a useful tool for preparing high density reproducible bilayer arrays to study small molecule–membrane interactions in a high-throughput manner.